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Establishment of cell lines stably expressing HNA‐1a, ‐1b, and ‐2a antigen with low background reactivity in flow cytometric analysis
Author(s) -
Yasui Kazuta,
Miyazaki Toru,
Matsuyama Nobuki,
Kojima Yoshitaka,
Furuta Rika A.,
Fujisawa Junichi,
Tani Yoshihiko,
Shibata Hirotoshi,
Sato Shinichirou,
Kato Toshiaki,
Ikeda Hisami,
Hirayama Fumiya
Publication year - 2007
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/j.1537-2995.2006.01139.x
Subject(s) - antibody , antigen , immunology , flow cytometry , neutropenia , cell culture , k562 cells , biology , microbiology and biotechnology , virology , medicine , leukemia , toxicity , genetics
BACKGROUND: Antibodies to neutrophil antigens have been implicated in neonatal alloimmune neutropenia, autoimmune neutropenia, and transfusion‐related acute lung injury. Most often, neutrophil‐specific antibodies are directed toward human neutrophil antigen (HNA)‐1 (Fcγ receptor 3b) and HNA‐2a (CD177) in these disorders. STUDY DESIGN AND METHODS: To detect the alloantibodies in the serum samples, a panel of cell lines was established in which the HNA‐1a, HNA‐1b (polymorphisms of HNA‐1), or HNA‐2a gene was transduced with a retrovirus vector to confer stable transgene expression in K562 cells that exhibited low background reactivity to human serum samples obtained from healthy donors in flow cytometric analysis. RESULTS: It was shown that several well‐characterized human serum samples containing antibodies against HNA‐1a, ‐1b, and ‐2a were unambiguously identified by the established panel cell lines and observed a lower background reactivity and longer shelf life of the K562 panel cell lines compared with isolated neutrophils, which have been used for the cell panel to identify antibodies against HNA in human serum samples. CONCLUSION: These results indicate that the K562 panel cell lines provide a good panel for detecting HNA‐reactive neutrophil antibodies in human serum samples.